Application of cannabidiol 2-2-(3-(butyl-3-alkyn-1-yl)-3H-bis (aziridine-3-yl) ethyl ester in treatment of type 2 diabetes mellitus

Abstract

The invention discloses an application of cannabidiol 2-2-(3-(butyl-3-alkyn-1-yl)-3H-bis (aziridine-3-yl) ethyl ester (hereinafter referred to as a2) in relieving symptoms of type 2 diabetes mellitus, and particularly relates to a compound shown in a formula I. The a2 has a reducing effect on blood sugar of mice suffering from type 2 diabetes mellitus, and can be used for preparing a medicine for relieving symptoms of type 2 diabetes mellitus. The oxidative stress of mice with type 2 diabetes can be relieved, and the insulin secretion of mouse islet beta cells can be promoted. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to the application of cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridine-3-yl)ethyl ester in preventing and treating diabetes. Background Art

[0002] Cannabidiol (CBD) is a major non-psychoactive drug extracted from the cannabis plant. Preclinical cell and rodent studies have shown that CBD may have neuroprotective, cardioprotective, and anti-inflammatory effects.

[0003] High-purity CBD is a white or light yellow crystal with a melting point of 66-67°C. It is almost insoluble in water, but easily soluble in organic solvents such as ethanol, methanol, ether, benzene, chloroform and petroleum ether. The ester solubility of CBD makes it very limited in drug development and finding accurate drug targets. Based on the structure of cannabidiol, designing and synthesizing cannabidiol derivatives is an important way to obtain more active compounds, which is of great significance for finding CBD targets and preparing new drugs.

[0004] Diabetes is a metabolic disease characterized by high blood sugar levels, which occurs when the pancreas can no longer produce insulin, or when the body cannot use the insulin produced by the pancreas well. Diabetes is mainly divided into type 1 diabetes, type 2 diabetes and gestational diabetes. Type 2 diabetes is the most common type, accounting for 90% of all types of diabetes. The "Guidelines for the Prevention and Treatment of Type 2 Diabetes in China (2020 Edition)" points out that according to the standards of the World Health Organization (WHO), the prevalence of diabetes in my country has reached 11.2%, of which more than 90% are patients with type 2 diabetes.

[0005] At present, the drugs for the treatment of type 2 diabetes have certain side effects to a greater or lesser extent. Among them, metformin, as the first-line drug for the treatment of type 2 diabetes, can relieve insulin resistance, but the most common adverse reactions of metformin are gastrointestinal reactions, which manifest as upper abdominal discomfort, abdominal pain, diarrhea, nausea, vomiting, bloating, fatigue, indigestion, etc. In addition, 10% of diabetic patients develop lactic acidosis after taking metformin. This causes some patients to give up due to intolerance to metformin. Therefore, it is very important to find safer and more effective drugs for the treatment of type 2 diabetes.

[0006] Designing and synthesizing cannabidiol derivatives based on the structure of cannabidiol is an important way to obtain more active compounds, which is of great significance for finding CBD targets and preparing new drugs.

[0007] Cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester is a cannabidiol derivative synthesized by us in the early stage. It has been verified that it can be used as a probe molecule for cannabidiol. Using click chemistry, we have used it to prove that cannabidiol is located in the mitochondria of cells and identified the target protein of cannabidiol. Summary of the invention

[0008] The inventors have found through in-depth research and creative work that cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridine-3-yl)ethyl ester can effectively treat and / or prevent type 2 diabetes, thereby providing the following invention:

[0009] One aspect of the present invention is the use of any one of the following (1) to (3) in the preparation of a medicament for treating and / or preventing type 2 diabetes:

[0010] (1) cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt, ester or solvate thereof;

[0011] (2) a mixture containing cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester;

[0012] (3) A pharmaceutical composition comprising cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt or ester or solvate thereof, and one or more pharmaceutically acceptable excipients;

[0013] The type 2 diabetes mellitus model mice were induced by a high-fat diet plus multiple injections of low-dose streptozotocin.

[0014] Cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester is a cannabidiol derivative, and its structural formula is shown in the following formula I:

[0015]

[0016] The cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester, i.e., the compound of formula I, can be synthesized by existing technology. After synthesis, the product can be separated, purified, and collected by silica gel plate.

[0017] Use of any one of the following (1) to (3) in the preparation of a drug for promoting insulin secretion in β cells:

[0018] (1) cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt, ester or solvate thereof;

[0019] (2) a mixture containing cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester;

[0020] (3) A pharmaceutical composition comprising cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt or ester or solvate thereof, and one or more pharmaceutically acceptable excipients. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 .Chemical structural formula of cannabidiol and cannabidiol derivative a2;

[0022] Figure 2 :The test results of cannabidiol and cannabidiol derivative a2 on fasting blood glucose in type 2 diabetes model mice;

[0023] Figure 3 :The test results of cannabidiol and cannabidiol derivative a2 on glucose tolerance in type 2 diabetes model mice;

[0024] Figure 4 :The test results of cannabidiol and cannabidiol derivative a2 on insulin sensitivity in type 2 diabetes model mice;

[0025] Figure 5 :The results of the detection of the oxidative stress marker glutathione content in the serum of type 2 diabetes model mice by cannabidiol and cannabidiol derivative a2;

[0026] Figure 6 :The results of the detection of cannabidiol and cannabidiol derivative a2 on the content of malondialdehyde, a marker of oxidative stress, in the serum of type 2 diabetes model mice;

[0027] Figure 7 :The results of the detection of the oxidative stress marker superoxide dismutase in the serum of type 2 diabetes model mice by cannabidiol and cannabidiol derivative a2;

[0028] Figure 8 :The results of detecting the effects of cannabidiol and cannabidiol derivative a2 on glucose-stimulated insulin secretion in MIN6 cells. DETAILED DESCRIPTION

[0029] The following is a further description of the concept of the present invention and the technical effects produced in conjunction with specific embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention. The methods are conventional methods unless otherwise specified. The materials can be obtained from public commercial channels unless otherwise specified.

[0030] Example 1: Determination of the effect of cannabidiol derivative a2 on lowering blood sugar in type 2 diabetic mice

[0031] The experimental animals used were 4-week-old C57BL male mice. The mice were divided into four groups (control group, type 2 diabetes model group, cannabidiol treatment group, cannabidiol derivative a2 treatment group), with five mice in each group. The control group mice were fed with ordinary feed for eight weeks, and the other groups of mice were fed with high-fat feed for eight weeks. After the mice fasted for 10-12 hours in the eighth week, the control group mice were intraperitoneally injected with an equal amount of streptozotocin (STZ) solvent, and the other groups of mice were intraperitoneally injected with 50 mg / kg STZ. The injection was continued for 4 days, and the fasting blood glucose values ​​of the mice were tested one week later to see whether the induction was successful (fasting blood glucose was greater than 11.1 mmol / L for successful induction). After the type 2 diabetes model was successfully induced, each mouse in the cannabidiol treatment group was given 50 mg / kg cannabidiol by gavage every day, and each mouse in the cannabidiol derivative a2 treatment group was given 50 mg / kg cannabidiol derivative a2 by gavage every day. The drug was administered for eight consecutive weeks, and the fasting blood glucose of the mice was tested with blood glucose test strips after fasting for 10-12 hours every two weeks.

[0032] The experimental results are as follows Figure 2 shown.

[0033] Figure 2 This is the test result of cannabidiol and cannabidiol derivative a2 on fasting blood glucose of type 2 diabetes model mice. The results show that cannabidiol derivative a2 can reduce fasting blood glucose of type 2 diabetes model mice.

[0034] Example 2: Determination of the effect of cannabidiol derivative a2 on improving glucose tolerance and insulin tolerance in type 2 diabetic mice

[0035] Determination of glucose tolerance in each group of mice

[0036] In the sixth week of administration, the mice were fasted for 12-16 hours at night and tested the next morning. Fasting blood sugar was tested first, and then 20% glucose was injected into each group of mice according to the standard of 10μL per 1g body weight. From then on, the timing began and the blood sugar of the mice was tested at 30min, 60min, 90min, and 120min. After the experiment, feed was added to each group of mice.

[0037] The experimental results are as follows Figure 3 shown.

[0038] Figure 3 This is the test result of cannabidiol and cannabidiol derivative a2 on glucose tolerance in type 2 diabetes model mice. The results show that cannabidiol derivative a2 can alleviate glucose intolerance in type 2 diabetes model mice.

[0039] Determination of insulin sensitivity of mice in each group.

[0040] In the seventh week of administration, each group of mice was fasted for 5-7 hours in the morning and tested in the afternoon. First, the body weight was weighed and the initial blood sugar was tested. Then, each group of mice was injected with insulin at the standard of 0.75U per kilogram of body weight. From then on, the timing began and the blood sugar of the mice was tested at 30min, 60min, 90min, and 120min. After the experiment, feed was added to each group of mice.

[0041] The experimental results are as follows Figure 4 shown.

[0042] Figure 4 This is the test result of cannabidiol and cannabidiol derivative a2 on insulin sensitivity of type 2 diabetes model mice. The results show that cannabidiol derivative a2 can alleviate insulin resistance in type 2 diabetes model mice.

[0043] Example 3: Determination of the effect of cannabidiol derivative a2 on reducing oxidative stress in type 2 diabetic mice

[0044] In the eighth week of administration, each group of mice was fasted overnight for 12-16 hours, and blood was collected from the eyeballs. Serum was collected after standing still, and the levels of oxidative stress markers SOD, MDA, and GSH in serum were detected using a total superoxide dismutase (T-SOD) assay kit, a malondialdehyde (MDA) test kit, and a reduced glutathione (GSH) assay kit, respectively.

[0045] The experimental results are as follows Figure 5 , as shown in Figures 6 and 7.

[0046] Figure 5 This is the result of the test of cannabidiol and cannabidiol derivative a2 on the content of glutathione, a marker of oxidative stress, in the serum of type 2 diabetes model mice.

[0047] Figure 6 This is the test result of cannabidiol and cannabidiol derivative a2 on the content of malondialdehyde, a marker of oxidative stress, in the serum of type 2 diabetes model mice.

[0048] Figure 7 This is the result of the test of cannabidiol and cannabidiol derivative a2 on the content of superoxide dismutase, a marker of oxidative stress, in the serum of type 2 diabetes model mice.

[0049] The results showed that cannabidiol derivative a2 can alleviate the oxidative stress level in type 2 diabetic mice.

[0050] Example 4: Determination of the effect of cannabidiol derivative a2 on insulin secretion stimulated by glucose in pancreatic β-cell line MIN6 cells

[0051] MIN6 cells were cultured at 1×10 6 / mL was plated in a six-well cell culture plate, and three groups (control group, cannabidiol treatment group, cannabidiol derivative a2 treatment group) were set up, with three replicates in each group. After the cells adhered to the wall, the cells were treated in groups. 2mL of complete medium was added to each well of the control group, 2mL of complete medium containing 10μM cannabidiol was added to each well of the cannabidiol treatment group, and 2mL of complete medium containing 30μM cannabidiol derivative a2 was added to each well of the cannabidiol derivative a2 treatment group. After 24h of treatment, the supernatant was discarded, and the cells were washed three times with PBS buffer. 1mL of sugar-free KRB buffer was added to each well and incubated at 37℃ for 1h. After 1h, the supernatant was discarded, and 1mL of KRB buffer containing 16.7mM glucose was added to each well and incubated at 37℃ for 1h. After 1h, the supernatant was collected, and 100μL of sugar-free KRB buffer was added after freeze-drying. The insulin concentration of each group was detected by radioimmunoassay. After the cells were collected, the protein concentration was detected by BSA method.

[0052] The experimental results are as follows Figure 8 shown.

[0053] Figure 8 The results of the test of cannabidiol and cannabidiol derivative a2 on glucose-stimulated insulin secretion of MIN6 cells. The results showed that treatment of MIN6 cells with cannabidiol and cannabidiol derivative a2 for 24 hours can significantly increase the glucose-stimulated insulin secretion of MIN6 cells.

[0054] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A cannabidiol derivative, characterized in that: The derivative is cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester, and the specific chemical structure is shown in Formula I:

2. Use of any one of the following (1) to (3) in the preparation of a medicament for treating and / or preventing type 2 diabetes: (1) cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt, ester or solvate thereof; (2) a mixture containing cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester; (3) A pharmaceutical composition comprising cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt or ester or solvate thereof, and one or more pharmaceutically acceptable excipients; The type 2 diabetes mellitus model mice were induced by a high-fat diet plus multiple injections of low-dose streptozotocin.

3. Use of any one of the following (1) to (3) in the preparation of a drug for promoting insulin secretion in β cells: (1) cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt, ester or solvate thereof; (2) a mixture containing cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester; (3) A pharmaceutical composition comprising cannabidiol-2-2-(3-(but-3-yn-1-yl)-3H-diaziridin-3-yl)ethyl ester or a pharmaceutically acceptable salt or ester or solvate thereof, and one or more pharmaceutically acceptable excipients.

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